Skip to main content

The In Situ Characterization of Membrane-Immobilized 2-D PAGE-Separated Proteins Using Ink-Jet Technology

  • Protocol
The Proteomics Protocols Handbook

Part of the book series: Springer Protocols Handbooks ((SPH))

  • 4115 Accesses

Abstract

A progression in technology development for proteomics research is occurring at an ever-increasing rate (1). The monitoring of the physiological changes of healthy and diseased tissues with linkage to the expression of the proteome is fast becoming a method to identify molecular disease targets for creating novel drugs, as well as providing data for basic research. Improvements in the preparation of protein samples and mass spectrometry (MS) equipment are leading to better identification and characterization of proteins (2,3). Advances in protocols for protein sample prefractionation, solubilization strategies, and two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) are oriented with the development of automated high-throughput proteomic analysis platforms (46).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Institutional subscriptions

References

  1. Lee, K. H. (2001) Proteomics: a technology-driven and technology-limited discovery science. Trends Biotechnol. 19, 217–222.

    Article  PubMed  CAS  Google Scholar 

  2. Hamdan, M. and Righetti, P. G. (2002) Modern strategies for protein quantification in proteome analysis: Advantages and limitations. Mass Spectrom. Rev. 21, 287–302.

    Article  PubMed  CAS  Google Scholar 

  3. Smith, R. D. (2002) Trends in mass spectrometry instrumentation for proteomics. Trends Biotechnol. 20, S3–S7.

    Article  PubMed  CAS  Google Scholar 

  4. Righetti, P. G., Castagna, A., and Herbert, B. (2001) Prefractionation techniques in proteome analysis. Anal. Chem. 73, 320A–326A.

    Article  PubMed  CAS  Google Scholar 

  5. Govorun, V. M., and Archakov, A. I. (2002) Proteomic technologies in modern biomedical science. Biochemistry 67, 1109–1123.

    PubMed  CAS  Google Scholar 

  6. Gygi, S. P., Corthals, G. L., Zhang, Y., Rochon, Y., and Aebersold, R. (2000) Evaluation of two-dimensional gel electrophoresis-based proteome analysis technology. Proc. Natl. Acad. Sci. USA 97, 9390–9395.

    Article  PubMed  CAS  Google Scholar 

  7. Wilson, D. S. and Nock, S. (2003) Recent developments in protein microarray technology. Angew. Chem. Int. Ed. Engl. 42, 494–500.

    Article  PubMed  CAS  Google Scholar 

  8. Lee, Y. S. and Mrksich, M. (2002) Protein chips: from concept to practice. Trends Biotechnol. 20, S14–18.

    Article  PubMed  CAS  Google Scholar 

  9. Zhu, H. and Snyder, M. (2003) Protein chip technology. Curr. Opin. Chem. Biol. 7, 55–63.

    Article  PubMed  CAS  Google Scholar 

  10. Sanders, G. H. and Manz, A. (2000) Chip-based Microsystems for genomic and proteomic analysis. Trends Anal. Chem. 19, 364–378.

    Article  CAS  Google Scholar 

  11. Figeys, D. and Pinto, D. (2001) Proteomics on a chip: promising developments. Electrophoresis 22, 208–216.

    Article  PubMed  CAS  Google Scholar 

  12. Candiano, G., Musante, L., Bruschi, M., et al. (2002) Two-dimensional maps in soft immobilized pH gradient gels: a new approach to the proteome of the Third Millennium. Electrophoresis 23, 292–297.

    Article  PubMed  CAS  Google Scholar 

  13. Gorg, A., Boguth, G., Kopf, A., Reil, G., Parlar, H., and Weiss, W. (2002) Sample prefractionation with Sephadex isoelectric focusing prior to narrow pH range two-dimensional gels. Proteomics 2, 1652–1657.

    Article  PubMed  CAS  Google Scholar 

  14. Westbrook, J. A., Yan, J. X., Wait, R., Welson, S. Y., and Dunn, M. J. (2001) Zooming-in on the proteome: very narrow-range immobilized pH gradients reveal more protein species and isoforms. Electrophoresis 22, 2865–2871.

    Article  PubMed  CAS  Google Scholar 

  15. Pedersen, S. K., Harry, J. L., Sebastian, L., et al. (2003) Unseen proteome: mining below the tip of the iceberg to find low abundance and membrane proteins. J. Proteome Res. 2, 303–311.

    Article  PubMed  CAS  Google Scholar 

  16. Sloane, A. J., Duff, J. L., Wilson, N. L., et al. (2002) High throughput peptide mass fingerprinting and protein macroarray analysis using chemical printing strategies. Mol. Cell. Proteomics 1, 490–499.

    Article  PubMed  CAS  Google Scholar 

  17. Henzel, W. J., Billeci, T. M., Stults, J. T., Wong, S. C., Grimley, C., and Watanabe, C. (1993) Identifying proteins from two-dimensional gels by molecular mass searching of peptide fragments in protein sequence databases. Proc. Natl. Acad. Sci. USA 90, 5011–5015.

    Article  PubMed  CAS  Google Scholar 

  18. Wilson, N. L., Schulz, B. L., Karlsson, N. G., and Packer, N. H. (2002) Sequential analysis of N-and O-linked glycosylation of 2-D PAGE separated glycoproteins. J. Proteome Res. 1, 521–529.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2005 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Cooley, P.W., Joss, J.L., Hopwood, F.G., Wilson, N.L., Gooley, A.A. (2005). The In Situ Characterization of Membrane-Immobilized 2-D PAGE-Separated Proteins Using Ink-Jet Technology. In: Walker, J.M. (eds) The Proteomics Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1385/1-59259-890-0:341

Download citation

  • DOI: https://doi.org/10.1385/1-59259-890-0:341

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-343-5

  • Online ISBN: 978-1-59259-890-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics